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Mapping of psoralen cross-linked nucleotides in RNA

机译:在RNA中的性交交联核苷酸的测绘

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A method is described for using the cross-linking reagent 4′-(hydroxy-methyl)-4,5′,8-trimethylpsoralen (HMT) to map bsss paired regions and higher-order structure within RNA molecules. Applying this method to yeast tRNAPhe, we have specifically identified cross-links within the acceptor stem between U6 X U68 in the D-stem between C11 X C25 and in the Tψ-stem between U50 X C63 and U52 X C63. We have also identified a unique cross-link between U8 X C48 which are trans pyrimidines in the core region due to tertiary interactions between U8:A14 and C48:G15 The precise point of cross-linking was deduced in every case by using purine-specific U2 ribo-nuclease along with cytidine-specific CL3 ribonuclease which will anomalously cleave after photoreversed pyrimidines. The ability to map the precise point of cross-linking should prove invaluable in identifying nucleotides in close proximity within the tertiary structure of other RNA molecules.
机译:描述用于使用交联试剂4' - (羟甲基)-4,5',8-三甲基乙醚(HMT)来映射BSSS成对区域和RNA分子内的高阶结构。将该方法应用于酵母TRNA pHE ,我们在D-step中的U 6 xu 68 之间的受体杆内的交联在C 11 xc 25之间,在u 50 xc 63之间的t 1-shep中, 63 和u 52 xc 63 。我们还确定了U 8 XC 48 之间的独特交叉链路,其由于U 8 之间的三级相互作用,在核心区域中是核心区域中的反式嘧啶: 14 和c 48 :g 15 通过使用嘌呤特定的u 2 核糖核酸酶以及细胞苷特异性Cl3核糖核酸酶,其在光探测嘧啶后会大致地切割。映射精确的交联点的能力应该证明在鉴定在其他RNA分子的三级结构内的近距离接近的核苷酸中的核苷酸。

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